摘要
目的探讨卵巢癌细胞顺铂(DDP)耐药后的转录组差异,并基于此筛选找寻潜在的拮抗剂。方法以A2780细胞为研究对象,构建DDP耐药细胞A2780-DDP;通过转录组学测序分析,找寻DDP耐药的关键因素,并以实时荧光定量PCR(qPCR)和Western blot实验进行验证;通过小分子抑制剂筛选,采用CCK-8细胞活力检测的方法找寻潜在拮抗剂。结果成功构建了A2780-DDP细胞株,发现耐药后细胞增殖无差异,但细胞侵袭和迁移能力增强;通过转录组学测序分析,发现ITGB7、Akt等可能是DDP耐药的关键基因,且qPCR和Western blot验证发现两者在A2780-DDP细胞株中存在高表达。CCK-8结果显示,雷公藤内酯醇(TPL)、奥拉帕尼等在A2780-DDP细胞株中具有较好的抑制效果。结论ITGB7/Akt通路在DDP耐药中发挥重要作用,TPL、奥拉帕尼等潜在的DDP耐药拮抗剂可为卵巢癌治疗提供新思路。
Objective To investigate the transcriptome differences of ovarian cancer cells after cisplatin(DDP)resistance,and to find potential antagonists based on this screening.Methods DDP-resistant cell line A2780-DDP was constructed with A2780 cells as the research object.Through transcriptome sequencing analysis,the key factors of DDP resistance were found and verified by quantitative real-time PCR(qPCR)and Western blot experiments.Through the screening of small molecule inhibitors,CCK-8 cell viability assay was used to find potential antagonists.Results A2780-DDP were successfully constructed,and it was found that there was no difference in cell proliferation after drug resistance,but the ability of cell invasion and migration was enhanced.Through transcriptome sequencing analysis,it was found that ITGB7 and Akt may be the key genes of A2780-DDP,and qPCR and Western blot showed that they were highly expressed in A2780-DDP.CCK-8 results showed that triptolide(TPL)and Olaparib had good inhibitory effects in DDP-resistant cell lines.Conclusion The ITGB7/Akt pathway plays an important role in DDP resistance,and potential DDP resistance antagonists such as TPL can provide new ideas for the treatment of ovarian cancer.
作者
王晓晓
杜佳慧
李莉蓉
郭伟强
刘松柏
WANG Xiaoxiao;DU Jiahui;LI Lirong;GUO Weiqiang;LIU Songbai(Suzhou Key Laboratory of Medical Biotechnology,Suzhou Vocational Health College,Suzhou,Jiangsu 215009,China;Suzhou University of Science and Technology,Suzhou,Jiangsu 215009,China)
出处
《重庆医学》
CAS
2024年第4期481-486,共6页
Chongqing medicine
基金
江苏省“六大人才高峰”高层次人才项目(YY-170)
江苏省333人才项目(BRA2019329)
江苏省苏州市科技局民生科技医疗卫生科技创新关键技术攻关项目(SKY2021029)
江苏高校“青蓝工程”中青年学术带头人(2021)
苏州卫生职业技术学院自然科学研究项目(szwzy201904)
2021年度省高校优秀科技创新团队(血液肿瘤新型标志物的鉴定及应用)。
关键词
顺铂
小分子抑制剂
卵巢癌
RNA-SEQ
耐药机制
cisplatin
small molecule inhibitor
ovarian cancer
RNA-seq
drug resistance mechanism
